linear quadrupole ion trap ltq orbitrap mass spectrometer  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99

    Structured Review

    Thermo Fisher linear quadrupole ion trap ltq orbitrap mass spectrometer
    Workflow to identify lipid raft-associated proteins. Stable isotopic labeling of amino acids in cell culture (SILAC) labeled SH-SY5Y cells left untreated or exposed to 400 nM okadaic acid for 50 min before mixed in a ratio 1:1 in 0.7% Triton X-100/MES buffer. After centrifugation in a sucrose gradient, 10 fractions of 0.5 mL were collected. The lipid raft fractions were identified by Western blotting using flotillin as a lipid raft marker. Fractions containing flotillin were pooled prior to removal of lipids by chloroform/methanol extraction, followed by acetone precipitation. Proteins were separated by SDS-PAGE. Proteins were in-gel digested with trypsin, and peptides were purified either with (1) C18 or (2) IMAC/TiO 2 and analyzed on <t>LTQ-Orbitrap.</t> Proteins were identified with Mascot (ver. 2.2) and quantified using MaxQuant (ver. 1.13).
    Linear Quadrupole Ion Trap Ltq Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/linear quadrupole ion trap ltq orbitrap mass spectrometer/product/Thermo Fisher
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    linear quadrupole ion trap ltq orbitrap mass spectrometer - by Bioz Stars, 2020-07
    99/100 stars

    Related Products / Commonly Used Together

    ultimate 3000 nano lc system
    lipid raft proteome mapping

    Images

    1) Product Images from "Identification of Dynamic Changes in Proteins Associated with the Cellular Cytoskeleton after Exposure to Okadaic Acid"

    Article Title: Identification of Dynamic Changes in Proteins Associated with the Cellular Cytoskeleton after Exposure to Okadaic Acid

    Journal: Marine Drugs

    doi: 10.3390/md11061763

    Workflow to identify lipid raft-associated proteins. Stable isotopic labeling of amino acids in cell culture (SILAC) labeled SH-SY5Y cells left untreated or exposed to 400 nM okadaic acid for 50 min before mixed in a ratio 1:1 in 0.7% Triton X-100/MES buffer. After centrifugation in a sucrose gradient, 10 fractions of 0.5 mL were collected. The lipid raft fractions were identified by Western blotting using flotillin as a lipid raft marker. Fractions containing flotillin were pooled prior to removal of lipids by chloroform/methanol extraction, followed by acetone precipitation. Proteins were separated by SDS-PAGE. Proteins were in-gel digested with trypsin, and peptides were purified either with (1) C18 or (2) IMAC/TiO 2 and analyzed on LTQ-Orbitrap. Proteins were identified with Mascot (ver. 2.2) and quantified using MaxQuant (ver. 1.13).
    Figure Legend Snippet: Workflow to identify lipid raft-associated proteins. Stable isotopic labeling of amino acids in cell culture (SILAC) labeled SH-SY5Y cells left untreated or exposed to 400 nM okadaic acid for 50 min before mixed in a ratio 1:1 in 0.7% Triton X-100/MES buffer. After centrifugation in a sucrose gradient, 10 fractions of 0.5 mL were collected. The lipid raft fractions were identified by Western blotting using flotillin as a lipid raft marker. Fractions containing flotillin were pooled prior to removal of lipids by chloroform/methanol extraction, followed by acetone precipitation. Proteins were separated by SDS-PAGE. Proteins were in-gel digested with trypsin, and peptides were purified either with (1) C18 or (2) IMAC/TiO 2 and analyzed on LTQ-Orbitrap. Proteins were identified with Mascot (ver. 2.2) and quantified using MaxQuant (ver. 1.13).

    Techniques Used: Isotopic Labeling, Cell Culture, Labeling, Centrifugation, Western Blot, Marker, SDS Page, Purification

    Related Articles

    High Performance Liquid Chromatography:

    Article Title: 14-3-3 binding to LRRK2 is disrupted by multiple Parkinson's disease-associated mutations and regulates cytoplasmic localization
    Article Snippet: .. The HPLC system was coupled to a linear ion-trap–orbitrap hybrid mass spectrometer (LTQ-Orbitrap XL, Thermo Fisher Scientific) via a nanoelectrospray ion source (Proxeon Biosystems) fitted with a 5 cm Picotip FS360-20-10 emitter. ..

    Article Title: Quantitative Proteomic Discovery of Dynamic Epigenome Changes that Control Human Cytomegalovirus (HCMV) Infection *
    Article Snippet: .. The HPLC effluent was directly coupled to an LTQ-Orbitrap XL hybrid mass spectrometer (ThermoFisher Scientific, San Jose, CA) with a resolution of 30,000 for full MS followed by seven data-dependent and targeted MS/MS acquisitions. .. To quantify RNA levels of DOT1L, total cellular RNA was isolated using Tri-Reagent (Sigma). cDNA was generated using 1 μg of RNA with TaqMan Reverse Transcription Reagents (Roche), and equal volumes of cDNA was utilized for quantitative PCR (qPCR).

    Quantitation Assay:

    Article Title: Investigation of Age-Specific Behavioral and Proteomic Changes in an Animal Model of Chronic Ethanol Exposure
    Article Snippet: .. Mass spectrometric analysis for iTRAQ-based quantitation is carried out on a hybrid linear ion trap-Orbitrap mass spectrometer (LTQ Orbitrap XL, Thermo Fisher Scientific) ( see Note ). .. Several data analysis software packages can be used for processing of mass spectrometric data for relative protein quantitation by either spectral counting or iTRAQ.

    Mass Spectrometry:

    Article Title: O-Glycosylation Modulates Proprotein Convertase Activation of Angiopoietin-like Protein 3
    Article Snippet: .. The product of ANGPTL3 dodecapeptide with GalNAc-T2 was characterized by electrospray ionization-linear ion trap-Fourier transform mass spectrometry in an LTQ-Orbitrap XL hybrid spectrometer (Thermo-Scientific, Bremen, Germany), equipped with electron transfer dissociation (ETD) for peptide sequence analysis by MS/MS (MS2 ) with retention of glycan site-specific fragments. .. The sample was dissolved in methanol/water (1:1) containing 1% formic acid and introduced by direct infusion via a TriVersa NanoMate ESI-Chip interface (Advion BioSystems, Ithaca, NY), at a flow rate of ∼100 nl/min and 1.4 kV spray voltage.

    Article Title: Investigation of Age-Specific Behavioral and Proteomic Changes in an Animal Model of Chronic Ethanol Exposure
    Article Snippet: .. Mass spectrometric analysis for iTRAQ-based quantitation is carried out on a hybrid linear ion trap-Orbitrap mass spectrometer (LTQ Orbitrap XL, Thermo Fisher Scientific) ( see Note ). .. Several data analysis software packages can be used for processing of mass spectrometric data for relative protein quantitation by either spectral counting or iTRAQ.

    Article Title: Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity *
    Article Snippet: .. Characterization of O-Glycosylation Sites Products of O -glycosylation reactions were characterized by electrospray ionization-linear ion trap-Fourier transform mass spectrometry (ESI-LIT-FT-MS) in an LTQ-Orbitrap XL hybrid spectrometer (Thermo-Scientific) equipped for both high energy collision-induced dissociation ( ) in an external collision cell , and electron transfer dissociation (ETD) ( ) for peptide sequence analysis by MS/MS (MS2 ) with retention of glycan site-specific fragments. .. Samples were dissolved in methanol/water (1:1) containing 1% formic acid and introduced by direct infusion via a TriVersa NanoMate ESI-Chip interface (Advion BioSystems) at a flow rate of ∼100 nl/min and 1.4 kV spray voltage.

    Article Title: 14-3-3 binding to LRRK2 is disrupted by multiple Parkinson's disease-associated mutations and regulates cytoplasmic localization
    Article Snippet: .. The HPLC system was coupled to a linear ion-trap–orbitrap hybrid mass spectrometer (LTQ-Orbitrap XL, Thermo Fisher Scientific) via a nanoelectrospray ion source (Proxeon Biosystems) fitted with a 5 cm Picotip FS360-20-10 emitter. ..

    Article Title: An Internal Standard-Assisted Synthesis and Degradation Proteomic Approach Reveals the Potential Linkage between VPS4B Depletion and Activation of Fatty Acid β-Oxidation in Breast Cancer Cells
    Article Snippet: .. LC-MS/MS Analysis Peptides were dissolved in 0.5% acetic acid (solvent A) and separated on a 10 cm reverse-phase PicoFrit spray tip (New Objective, Woburn, MA) packed in house with sub-2 μ m C18 resin (Prospereon Life Science, IL), using a nanoflow Xtreme simple liquid chromatography system (Microtech/CVC) coupled to a hybrid linear ion trap-Orbitrap mass spectrometer (LTQ Orbitrap, Thermo Scientific). .. Peptides were loaded onto the column with solvent A at a flow rate of 0.6 μ L/min and eluted with a 180 min linear gradient at a flow rate of 0.2 μ L/min.

    Article Title: Quantitative HPLC-MS analysis of nucleotide sugars in plant cells following off-line SPE sample preparation
    Article Snippet: .. Fragmentation experiments were performed on a linear ion trap–Orbitrap mass spectrometer (Model LTQ Orbitrap XL, Thermo Fisher Scientific). .. Establishing stable retention times for UDP-sugars on PGC Figure illustrates the effect of the electrical potential applied to the ESI source on chromatographic retention of UDP-sugars on PGC.

    Article Title: Quantitative Proteomic Discovery of Dynamic Epigenome Changes that Control Human Cytomegalovirus (HCMV) Infection *
    Article Snippet: .. The HPLC effluent was directly coupled to an LTQ-Orbitrap XL hybrid mass spectrometer (ThermoFisher Scientific, San Jose, CA) with a resolution of 30,000 for full MS followed by seven data-dependent and targeted MS/MS acquisitions. .. To quantify RNA levels of DOT1L, total cellular RNA was isolated using Tri-Reagent (Sigma). cDNA was generated using 1 μg of RNA with TaqMan Reverse Transcription Reagents (Roche), and equal volumes of cDNA was utilized for quantitative PCR (qPCR).

    Tandem Mass Spectroscopy:

    Article Title: O-Glycosylation Modulates Proprotein Convertase Activation of Angiopoietin-like Protein 3
    Article Snippet: .. The product of ANGPTL3 dodecapeptide with GalNAc-T2 was characterized by electrospray ionization-linear ion trap-Fourier transform mass spectrometry in an LTQ-Orbitrap XL hybrid spectrometer (Thermo-Scientific, Bremen, Germany), equipped with electron transfer dissociation (ETD) for peptide sequence analysis by MS/MS (MS2 ) with retention of glycan site-specific fragments. .. The sample was dissolved in methanol/water (1:1) containing 1% formic acid and introduced by direct infusion via a TriVersa NanoMate ESI-Chip interface (Advion BioSystems, Ithaca, NY), at a flow rate of ∼100 nl/min and 1.4 kV spray voltage.

    Article Title: Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity *
    Article Snippet: .. Characterization of O-Glycosylation Sites Products of O -glycosylation reactions were characterized by electrospray ionization-linear ion trap-Fourier transform mass spectrometry (ESI-LIT-FT-MS) in an LTQ-Orbitrap XL hybrid spectrometer (Thermo-Scientific) equipped for both high energy collision-induced dissociation ( ) in an external collision cell , and electron transfer dissociation (ETD) ( ) for peptide sequence analysis by MS/MS (MS2 ) with retention of glycan site-specific fragments. .. Samples were dissolved in methanol/water (1:1) containing 1% formic acid and introduced by direct infusion via a TriVersa NanoMate ESI-Chip interface (Advion BioSystems) at a flow rate of ∼100 nl/min and 1.4 kV spray voltage.

    Article Title: Quantitative Proteomic Discovery of Dynamic Epigenome Changes that Control Human Cytomegalovirus (HCMV) Infection *
    Article Snippet: .. The HPLC effluent was directly coupled to an LTQ-Orbitrap XL hybrid mass spectrometer (ThermoFisher Scientific, San Jose, CA) with a resolution of 30,000 for full MS followed by seven data-dependent and targeted MS/MS acquisitions. .. To quantify RNA levels of DOT1L, total cellular RNA was isolated using Tri-Reagent (Sigma). cDNA was generated using 1 μg of RNA with TaqMan Reverse Transcription Reagents (Roche), and equal volumes of cDNA was utilized for quantitative PCR (qPCR).

    Sequencing:

    Article Title: O-Glycosylation Modulates Proprotein Convertase Activation of Angiopoietin-like Protein 3
    Article Snippet: .. The product of ANGPTL3 dodecapeptide with GalNAc-T2 was characterized by electrospray ionization-linear ion trap-Fourier transform mass spectrometry in an LTQ-Orbitrap XL hybrid spectrometer (Thermo-Scientific, Bremen, Germany), equipped with electron transfer dissociation (ETD) for peptide sequence analysis by MS/MS (MS2 ) with retention of glycan site-specific fragments. .. The sample was dissolved in methanol/water (1:1) containing 1% formic acid and introduced by direct infusion via a TriVersa NanoMate ESI-Chip interface (Advion BioSystems, Ithaca, NY), at a flow rate of ∼100 nl/min and 1.4 kV spray voltage.

    Article Title: Lectin Domains of Polypeptide GalNAc Transferases Exhibit Glycopeptide Binding Specificity *
    Article Snippet: .. Characterization of O-Glycosylation Sites Products of O -glycosylation reactions were characterized by electrospray ionization-linear ion trap-Fourier transform mass spectrometry (ESI-LIT-FT-MS) in an LTQ-Orbitrap XL hybrid spectrometer (Thermo-Scientific) equipped for both high energy collision-induced dissociation ( ) in an external collision cell , and electron transfer dissociation (ETD) ( ) for peptide sequence analysis by MS/MS (MS2 ) with retention of glycan site-specific fragments. .. Samples were dissolved in methanol/water (1:1) containing 1% formic acid and introduced by direct infusion via a TriVersa NanoMate ESI-Chip interface (Advion BioSystems) at a flow rate of ∼100 nl/min and 1.4 kV spray voltage.

    Liquid Chromatography:

    Article Title: An Internal Standard-Assisted Synthesis and Degradation Proteomic Approach Reveals the Potential Linkage between VPS4B Depletion and Activation of Fatty Acid β-Oxidation in Breast Cancer Cells
    Article Snippet: .. LC-MS/MS Analysis Peptides were dissolved in 0.5% acetic acid (solvent A) and separated on a 10 cm reverse-phase PicoFrit spray tip (New Objective, Woburn, MA) packed in house with sub-2 μ m C18 resin (Prospereon Life Science, IL), using a nanoflow Xtreme simple liquid chromatography system (Microtech/CVC) coupled to a hybrid linear ion trap-Orbitrap mass spectrometer (LTQ Orbitrap, Thermo Scientific). .. Peptides were loaded onto the column with solvent A at a flow rate of 0.6 μ L/min and eluted with a 180 min linear gradient at a flow rate of 0.2 μ L/min.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher hybrid linear quadrupole ion trap orbitrap mass spectrometer
    (a) CID on the <t>LTQ-Orbitrap</t> of the crosslinked homodimer synthetic peptide (Ac-QIGKGVAR) results in the same two major cleavage events observed in MALDI at the intrinsic positive charges. (b) MS 3 of the fragment without an intrinsic positive charge produces
    Hybrid Linear Quadrupole Ion Trap Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hybrid linear quadrupole ion trap orbitrap mass spectrometer/product/Thermo Fisher
    Average 99 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    hybrid linear quadrupole ion trap orbitrap mass spectrometer - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    85
    Thermo Fisher linear quadrupole ion trap ltq orbitrap mass spectrometer
    Workflow to identify lipid raft-associated proteins. Stable isotopic labeling of amino acids in cell culture (SILAC) labeled SH-SY5Y cells left untreated or exposed to 400 nM okadaic acid for 50 min before mixed in a ratio 1:1 in 0.7% Triton X-100/MES buffer. After centrifugation in a sucrose gradient, 10 fractions of 0.5 mL were collected. The lipid raft fractions were identified by Western blotting using flotillin as a lipid raft marker. Fractions containing flotillin were pooled prior to removal of lipids by chloroform/methanol extraction, followed by acetone precipitation. Proteins were separated by SDS-PAGE. Proteins were in-gel digested with trypsin, and peptides were purified either with (1) C18 or (2) IMAC/TiO 2 and analyzed on <t>LTQ-Orbitrap.</t> Proteins were identified with Mascot (ver. 2.2) and quantified using MaxQuant (ver. 1.13).
    Linear Quadrupole Ion Trap Ltq Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/linear quadrupole ion trap ltq orbitrap mass spectrometer/product/Thermo Fisher
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    linear quadrupole ion trap ltq orbitrap mass spectrometer - by Bioz Stars, 2020-07
    85/100 stars
      Buy from Supplier

    99
    Thermo Fisher hybrid dual cell quadrupole linear ion trap orbitrap mass spectrometer
    Solid phase extraction-based strong cation exchange (A) Shown is the solution charge state distribution of 38,487 phosphorylated and unphosphorylated peptides from lysC digested yeast whole-cell extract subjected to the described SPE/SCX/IMAC enrichment protocol analyzed by LC-MS/MS on a LTQ <t>Orbitrap</t> Velos. (B) The peptide overlap between SCX fractions is shown as the fraction of all peptides that were identified in 1, 2, 3, or more different fractions. (C) A comparison of the solution charge states of the identified phosphopeptides from trypsin and lysC digested yeast whole cell extracts.
    Hybrid Dual Cell Quadrupole Linear Ion Trap Orbitrap Mass Spectrometer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hybrid dual cell quadrupole linear ion trap orbitrap mass spectrometer/product/Thermo Fisher
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    hybrid dual cell quadrupole linear ion trap orbitrap mass spectrometer - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    (a) CID on the LTQ-Orbitrap of the crosslinked homodimer synthetic peptide (Ac-QIGKGVAR) results in the same two major cleavage events observed in MALDI at the intrinsic positive charges. (b) MS 3 of the fragment without an intrinsic positive charge produces

    Journal: Journal of the American Society for Mass Spectrometry

    Article Title: Quaternary Diamines as Mass Spectrometry Cleavable Crosslinkers for Protein Interactions

    doi: 10.1007/s13361-011-0288-4

    Figure Lengend Snippet: (a) CID on the LTQ-Orbitrap of the crosslinked homodimer synthetic peptide (Ac-QIGKGVAR) results in the same two major cleavage events observed in MALDI at the intrinsic positive charges. (b) MS 3 of the fragment without an intrinsic positive charge produces

    Article Snippet: The crosslinked standard peptides were analyzed with a hybrid linear quadrupole ion trap-Orbitrap mass spectrometer (ThermoFisher Scientific, Inc. model LTQ-Orbitrap XL; San Jose, CA, USA).

    Techniques: Mass Spectrometry

    Workflow to identify lipid raft-associated proteins. Stable isotopic labeling of amino acids in cell culture (SILAC) labeled SH-SY5Y cells left untreated or exposed to 400 nM okadaic acid for 50 min before mixed in a ratio 1:1 in 0.7% Triton X-100/MES buffer. After centrifugation in a sucrose gradient, 10 fractions of 0.5 mL were collected. The lipid raft fractions were identified by Western blotting using flotillin as a lipid raft marker. Fractions containing flotillin were pooled prior to removal of lipids by chloroform/methanol extraction, followed by acetone precipitation. Proteins were separated by SDS-PAGE. Proteins were in-gel digested with trypsin, and peptides were purified either with (1) C18 or (2) IMAC/TiO 2 and analyzed on LTQ-Orbitrap. Proteins were identified with Mascot (ver. 2.2) and quantified using MaxQuant (ver. 1.13).

    Journal: Marine Drugs

    Article Title: Identification of Dynamic Changes in Proteins Associated with the Cellular Cytoskeleton after Exposure to Okadaic Acid

    doi: 10.3390/md11061763

    Figure Lengend Snippet: Workflow to identify lipid raft-associated proteins. Stable isotopic labeling of amino acids in cell culture (SILAC) labeled SH-SY5Y cells left untreated or exposed to 400 nM okadaic acid for 50 min before mixed in a ratio 1:1 in 0.7% Triton X-100/MES buffer. After centrifugation in a sucrose gradient, 10 fractions of 0.5 mL were collected. The lipid raft fractions were identified by Western blotting using flotillin as a lipid raft marker. Fractions containing flotillin were pooled prior to removal of lipids by chloroform/methanol extraction, followed by acetone precipitation. Proteins were separated by SDS-PAGE. Proteins were in-gel digested with trypsin, and peptides were purified either with (1) C18 or (2) IMAC/TiO 2 and analyzed on LTQ-Orbitrap. Proteins were identified with Mascot (ver. 2.2) and quantified using MaxQuant (ver. 1.13).

    Article Snippet: For general lipid raft proteome mapping, the peptides were separated on a Dionex Ultimate 3000 nano-LC system (Sunnyvale, CA, USA) coupled to a linear quadrupole ion trap (LTQ-Orbitrap) mass spectrometer (Thermo Scientific).

    Techniques: Isotopic Labeling, Cell Culture, Labeling, Centrifugation, Western Blot, Marker, SDS Page, Purification

    Stem cell therapy-specific subproteome. ( A ): Comparison of stem cell-treated [ES(+)] versus untreated [ES(−)] left ventricular tissue extracts by two-dimensional (2D) electrophoresis. Differentially expressed spots isolated for identification by LTQ-Orbitrap mass spectrometric analysis are circled, and numbered on the ES(+) gel. Inset: Gel-to-gel reproducibility indicated by correlation of scatter plot for average normalized densitometric intensities of matching protein spots from ES(+) versus ES(−) gels. ( B ): Identities of the 61 proteins significantly altered by cell therapy are listed with their symbol (Swiss-Prot gene abbreviation) and spot numbers to locate corresponding 2D gel position(s) in panel ( A ). Mascot score, number of unique identified peptides, % sequence cov. (coverage), predicted M r and p I for each protein (following expected post-translational processing, for example, removal of a mitochondrial signal peptide), and fold change in ES(+) versus ES(−) are indicated. For proteins detected in more than one spot, maximum score and corresponding number of unique peptides are reported. Fold change was calculated as described in experimental procedures, and for proteins detected in both increasing and decreasing spots (*), both values are indicated. Abbreviations: ES, embryonic stem cells; Ox., oxidative; TCA cycle, tricarboxylic acid cycle; SAPK, stress activated protein kinase.

    Journal: Stem cells (Dayton, Ohio)

    Article Title: ATP-Sensitive K+ Channel-Deficient Dilated Cardiomyopathy Proteome Remodeled by Embryonic Stem Cell Therapy

    doi: 10.1002/stem.465

    Figure Lengend Snippet: Stem cell therapy-specific subproteome. ( A ): Comparison of stem cell-treated [ES(+)] versus untreated [ES(−)] left ventricular tissue extracts by two-dimensional (2D) electrophoresis. Differentially expressed spots isolated for identification by LTQ-Orbitrap mass spectrometric analysis are circled, and numbered on the ES(+) gel. Inset: Gel-to-gel reproducibility indicated by correlation of scatter plot for average normalized densitometric intensities of matching protein spots from ES(+) versus ES(−) gels. ( B ): Identities of the 61 proteins significantly altered by cell therapy are listed with their symbol (Swiss-Prot gene abbreviation) and spot numbers to locate corresponding 2D gel position(s) in panel ( A ). Mascot score, number of unique identified peptides, % sequence cov. (coverage), predicted M r and p I for each protein (following expected post-translational processing, for example, removal of a mitochondrial signal peptide), and fold change in ES(+) versus ES(−) are indicated. For proteins detected in more than one spot, maximum score and corresponding number of unique peptides are reported. Fold change was calculated as described in experimental procedures, and for proteins detected in both increasing and decreasing spots (*), both values are indicated. Abbreviations: ES, embryonic stem cells; Ox., oxidative; TCA cycle, tricarboxylic acid cycle; SAPK, stress activated protein kinase.

    Article Snippet: Chromatography was performed using 0.2% formic acid in solvent A (99% water, 1% acetonitrile) and B (80% acetonitrile, 5% isopropanol, 15% water), with peptides eluted over 30 minutes with a 5%–45% solvent B gradient using an Eksigent nanoHPLC system (MDS Sciex, Toronto, Canada) coupled to a linear ion trap quadrupole (LTQ)-Orbitrap mass spectrometer (Thermo Fisher Scientific, Barrington, IL).

    Techniques: Two-Dimensional Gel Electrophoresis, Isolation, Sequencing

    Solid phase extraction-based strong cation exchange (A) Shown is the solution charge state distribution of 38,487 phosphorylated and unphosphorylated peptides from lysC digested yeast whole-cell extract subjected to the described SPE/SCX/IMAC enrichment protocol analyzed by LC-MS/MS on a LTQ Orbitrap Velos. (B) The peptide overlap between SCX fractions is shown as the fraction of all peptides that were identified in 1, 2, 3, or more different fractions. (C) A comparison of the solution charge states of the identified phosphopeptides from trypsin and lysC digested yeast whole cell extracts.

    Journal: Methods (San Diego, Calif.)

    Article Title: A solid phase extraction-based platform for rapid phosphoproteomic analysis

    doi: 10.1016/j.ymeth.2011.03.008

    Figure Lengend Snippet: Solid phase extraction-based strong cation exchange (A) Shown is the solution charge state distribution of 38,487 phosphorylated and unphosphorylated peptides from lysC digested yeast whole-cell extract subjected to the described SPE/SCX/IMAC enrichment protocol analyzed by LC-MS/MS on a LTQ Orbitrap Velos. (B) The peptide overlap between SCX fractions is shown as the fraction of all peptides that were identified in 1, 2, 3, or more different fractions. (C) A comparison of the solution charge states of the identified phosphopeptides from trypsin and lysC digested yeast whole cell extracts.

    Article Snippet: Peptides were detected in a hybrid dual-cell quadrupole linear ion trap – orbitrap mass spectrometer (LTQ Orbitrap Velos, ThermoFisher) using a data-dependent Top20 method, or the single-cell linear ion trap – orbitrap mass spectrometer (LTQ Orbitrap Discovery, ThermoFisher) using a Top10 method.

    Techniques: Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry